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WO2025155882A1 - Composés dégradant/perturbant g9a/glp et procédés d'utilisation - Google Patents

Composés dégradant/perturbant g9a/glp et procédés d'utilisation

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Publication number
WO2025155882A1
WO2025155882A1 PCT/US2025/012138 US2025012138W WO2025155882A1 WO 2025155882 A1 WO2025155882 A1 WO 2025155882A1 US 2025012138 W US2025012138 W US 2025012138W WO 2025155882 A1 WO2025155882 A1 WO 2025155882A1
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Prior art keywords
optionally substituted
alkyl
membered
cycloalkyl
heterocyclyl
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Inventor
Jian Jin
Yan Xiong
Yulin HAN
Julia VELEZ
Hyerin Yim
Kabir, Md
Peiyi Yang
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Icahn School of Medicine at Mount Sinai
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Icahn School of Medicine at Mount Sinai
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • This disclosure relates to bivalent compounds (e.g., bi-functional small molecule compounds) which degrade and/or disrupt the G9a/GLP (G9a, also known as Vietnamese histonelysine N-methyl transferase 2 (EHMT2), GLP, also known as Vietnamese histone-lysine N- methyltransferase 1 (EHMT1) or G9a like protein ) compositions comprising one or more of the bivalent compounds, and to methods of use thereof for the treatment of G9a/GLP-mediated disease in a subject in need thereof.
  • the disclosure also relates to methods for designing such bivalent compounds.
  • G9a also known as Vietnamese histone-lysine N-methyltransferase 2 (EHMT2)
  • GLP G9a-like protein
  • EHMT1 are lysine methyltransferases that catalyze mono- and di-methylation of histone H3 lysine 9 (H3K9), transcriptionally repressive chromatin marks, and non-histone proteins.
  • G9a and GLP which share approximately 80% sequence homology in their catalytic SET domains, (Chang et al., 2009; Link et al., 2009) play important roles in diverse cellular processes including cell development, differentiation, and hypoxia response.
  • G9a/GLP axis When atypically expressed, the G9a/GLP axis promotes cancer progression, survival, and metastasis through several different mechanisms.
  • G9a/GLP In addition to G9a/GLP’s well-established histone methyltransferase activity, studies have demonstrated that G9a/GLP have non-canonical oncogenic functions, such as methylating nonhistone proteins including HDAC1, DNMT1, and p53.
  • G9a/GLP have non-catalytic oncogenic activity by functioning as a co-activator independent of its catalytic domain.
  • overexpression of G9a has been reported in several cancer types such as breast, (Tu et al., 2018; Wang et al., 2017) lung, (Chen et al., 2010; Sun et al., 2021) leukemia, (Lehnertz et al., 2014) bladder, (Cao et al., 2019; Segovia et al., 2019) colorectal, (Bergin et al., 20
  • G9a/GLP enzymatic inhibitors have been reported, (Kaniskan et al., 2018; Katayama et al., 2020; Kubicek et al., 2007; Liu et al., 2009; Milite et al., 2019; Moreira-Silva et al., 2022; San Jose-Eneriz et al., 2017; S Stamm et al., 2014; Takase et al., 2023; Vedadi et al., 2011; Xiong et al., 2017) including our in vivo G9a/GLP chemical probe UNC0642 (Liu et al., 2013) and G9a/GLP covalent inhibitor MS8511(Park et al., 2022).
  • G9a/GLP inhibitors effectively reduce the H3K9me2 mark in cells, they display limited cancer cell killing activity, potentially due to that they are unable to target G9a/GLP’s non-catalytic oncogenic functions.
  • G9a/GLP knockdown led to enhanced anti-proliferative effects through the induction of cell cycle arrest and apoptosis.
  • G9a/GLP degradation / disruption compounds bind and induce degradation of G9a/GLP, thus eliminating any scaffolding functions of G9a/GLP in addition to the kinase activity of G9a/GLP.
  • the G9a/GLP degraders disclosed herein are bivalent compounds, including a G9a/GLP ligand conjugated to a degradation / disruption tag via a linker.
  • the G9a/GLP degraders disclosed herein offer a novel mechanism for treating G9a/GLP- mediated diseases.
  • the present G9a/GLP degraders have the ability to target G9A/GLP for degradation, as opposed to merely inhibit the kinase activity of G9a/GLP.
  • this disclosure provides a method of treating G9a/GLP-mediated diseases, the method including administering one or more G9a/GLP degraders to a subject who has an G9a/GLP-mediated disease, the G9a/GLP degraders being bivalent compounds including a G9a/GLP ligand conjugated to a degradation/disruption tag via a linker.
  • the G9a/GLP-mediated disease can be a disease resulting from G9a/GLP expression.
  • the G9a/GLP-mediated disease can have elevated G9a/GLP expression relative to a wild-type tissue of the same species and tissue type.
  • Non-limiting examples of G9a/GLP-mediated diseases include breast cancer, lung cancer, prostate cancer, colon cancer, pancreatic cancer, bladder cancer, liver cancer, and cervical cancer.
  • the G9a/GLP-mediated cancer can include, e.g., a relapsed cancer.
  • the G9a/GLP- mediated cancer can, e.g., be refractory to one or more previous treatments.
  • the present disclosure relates generally to bivalent compounds (e.g., bi-functional small molecule compounds) which degrade and/or disrupt G9a/GLP, and to methods for the treatment of G9a/GLP-mediated cancer (i.e., a cancer which depends on G9a/GLP protein; or cancer having elevated levels of G9a/GLP, or G9a/GLP activity relative to a wild-type tissue of the same species and tissue type).
  • G9a/GLP-mediated cancer i.e., a cancer which depends on G9a/GLP protein; or cancer having elevated levels of G9a/GLP, or G9a/GLP activity relative to a wild-type tissue of the same species and tissue type.
  • the bivalent compounds of the present disclosure can be significantly more effective therapeutic agents than current G9a/GLP inhibitors, which inhibit the kinase activity of G9a/GLP, but do not affect G9a/GLP protein levels.
  • the present disclosure further provides methods for identifying G9a/GLP degraders/disruptors as described herein.
  • the present disclosure provides a bivalent compound including an G9a/GLP ligand conjugated to a degradation/disruption tag via a linker.
  • R 1 is H, halogen, cyano, NR a R b , C(O)NR a R b , C(O)R b , OR a , OR b , SR a , SR b or R S1 ; wherein R a and R b are independently selected from H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl optionally substituted with one or more halo, hydroxyl, oxo, amino, mono- or dialkylamino, or C 1 -C 6 alkoxyl; or R a and R b taken together with the nitrogen to which they are attached together form a 4-9 membered ring; wherein R S1 is selected from C 3 -C 8 cycloalkyl, phenyl, 4- to 12-membered heterocycloalkyl containing 1-4 heteroatoms selected from N, O, and S, spiro-fused 4- to 12-
  • X 2 is in each occurrence independently selected from N, or CH;
  • the FORMULA (1) can be FORMULA (1-a): or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer.
  • C is a ring structure selected from C 3 -C 10 cycloalkyl, C 6 -C 10 aryl, 4- to 12-membered heterocycloalkyl containing 1-4 heteroatoms selected from N, O, and S, or a 5- to 10-membered heteroaryl; n is selected from 0-12;
  • R 3b is selected from optionally substituted
  • C is selected from optionally substituted
  • R 1a is NH 2 . In some aspects of FORMULAE (1-a1), R 1a is In some aspects of FORMULAE (1-a1), R 1a is In some aspects of FORMULAE (I-b1), C is In some aspects of FORMULAE
  • D is In some aspects of
  • ring A is phenyl or pyridyl
  • R 1 is H or C 1 -C 4 alkyl
  • Each R 2 , R 3 , and R 4 independently is H, or C 1 -C 3 alkyl
  • R 5 is H, halogen, cyano, NR a R b , C(O)NR a R b , C(O)R b , OR a , OR b , SR a , SR b , R S1 or C 1 -C 3 alkylene, C 2 -C 4 alkenylene, C 2 -C 4 alkynylene linker optionally substituted with one or more of halo, cyano, hydroxyl, oxo, C 1 -C 3 alkyl or C 1 -C 3 alkoxyl; wherein R a and R b are independently selected from H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl optionally substituted with one or more halo, hydroxyl, oxo, amino, mono- or dialkylamino, or C 1 -C 6 alkoxyl; or R a and R b taken together with the nitrogen
  • V, W, and X are independently selected from CR 2 and N;
  • R 3 , and R 4 are independently selected from hydrogen, halogen, cyano, nitro, optionally substituted C 1 -C 6 alkyl, optionally substituted 3 to 6 membered carbocyclyl, and optionally substituted 4 to 6 membered heterocyclyl; or R 3 and R 4 together with the atom to which they are connected form a 3-6 membered carbocyclyl, or 4-6 membered heterocyclyl; and
  • R 3 is H, C(O)C 1 -C 8 alkyl, C(O)C 1 -C 8 alkoxyalkyl, C(O)C 1 -C 8 haloalkyl, C(O)C 1 -C 8 hydroxyalkyl, C(O)C 1 -C 8 aminoalkyl, C(O)C 1 -C 8 alkylaminoalkyl, C(O)C 3 -C 7 cycloalkyl, C(O)C 3 -C 7 heterocyclyl, C(O)C 2 -C 8 alkenyl, C(O)C 2 -C 8 alkynyl, C(O)OC 1 -C 8 alkoxyalkyl, C(O)OC 1 -C 8 haloalkyl, C(O)OC 1 -C 8 hydroxyalkyl, C(O)OC 1 -C 8 aminoalkyl, C(O)OC 1 -C 8 alkylaminoalkyl, C(O)
  • R 3 is hydrogen, optionally substituted C(O)C 1 -C 8 alkyl, optionally substituted C(O)C 1 - C 8 alkoxyC 1 -C 8 alkyl, optionally substituted C(O)C 1 -C 8 haloalkyl, optionally substituted C(O)C 1 - C 8 hydroxyalkyl, optionally substituted C(O)C 1 -C 8 aminoalkyl, optionally substituted C(O)C 1 - C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted C(O)C 3 -C 7 cycloalkyl, optionally substituted C(O)(3-7 membered heterocyclyl), optionally substituted C(O)C 2 -C 8 alkenyl, optionally substituted C(O)C 2 -C 8 alkynyl, optionally substituted C(O)OC 1 -C 8 alkoxyC 1 -C 8 alkyl, optionally substituted C(O)OC
  • R 4 and R 5 are independently selected from hydrogen, COR 6 , CO 2 R 6 , CONR 6 R 7 , SOR 6 , SO 2 R 6 , SO 2 NR 6 R 7 , optionally substituted C 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkoxyC 1 - C 8 alkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-8 membered cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, wherein
  • R 6 and R 7 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkoxy, optionally substituted C 1 -C 8 alkoxyC 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-8 membered cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or
  • Ar is selected from aryl and heteroaryl, each of which is optionally substituted with one or more substituents independently selected from F, Cl, CN, NO 2 , OR 8 , NR 8 R 9 , COR 8 , CO 2 R 8 , CONR 8 R 9 , SOR 8 , SO 2 R 8 , SO 2 NR 9 R 10 , NR 9 COR 10 , NR 8 C(O)NR 9 R 10 , NR 9 SOR 10 , NR 9 SO 2 R 10 , optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxyalkyl, optionally substituted C 1 -C 6 haloalkyl, optionally substituted C 1 -C 6 hydroxyalkyl, optionally substituted C 1 - C 6 alkylaminoC 1 -C 6 alkyl, optionally substituted C 3 -C 7 cycloalkyl, optionally substituted 3-7 membered heterocyclyl, optionally substituted C 2
  • degradation/disruption tags include a moiety according to FORMULA
  • V, W, X, and Z are independently selected from CR 4 and N;
  • R 1 , R 2 , R 3 , and R 4 are independently hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxyalkyl, C 1 -C 8 haloalkyl, C 1 -C 8 hydroxyalkyl, C 3 -C 7 cycloalkyl, C 3 -C 7 heterocyclyl, C 2 -C 8 alkenyl, or C 2 -C 8 alkynyl.
  • degradation/disruption tags include a moiety according to FORMULA
  • R 4 and R 5 are independently selected from hydrogen, COR 6 , CO 2 R 6 , CONR 6 R 7 , SOR 6 , SO 2 R 6 , SO 2 NR 6 R 7 , optionally substituted C 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkoxyC 1 - C 8 alkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted aryl-C 1 -C 8 alkyl, optionally substituted 3-8 membered cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, wherein
  • R 6 and R 7 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkoxyC 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-8 membered cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or
  • the degradation/disruption tag can be, for example, pomalidomide (Fischer et al., 2014), thalidomide (Fischer et al., 2014), lenalidomide (Fischer et al, 2014), VH032 (Galdeano et al., 2014; Maniaci et al., 2017), adamantine (Xie et al., 2014), 1 -((4, 4, 5,5,5- pentafluoropentyl)sulfinyl)nonane (E.Wakeling, 1995), nutlin-3a (Vassilev et al., 2004), RG7112 (Vu et al., 2013), RG7338, AMG 232 (Sun et al., 2014), AA-115 (Aguilar et al., 2017), bestatin (Hiroyuki Suda et al., 1976), MV1 (Varfolomeev
  • the linker can be a moiety of: wherein A, W and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R’-R”, R’COR ”, R’CO 2 R ”, R’C(O)N(R 1 )R”, R'C(S)N(R 1 )R”, R'OR ”, ROC(O)R”, R'OC(O)OR” , R OCON(R’)R” , R'SR”, R’SOR”, R'SO 2 R”, R'SO 2 N(R 1 )R”, R’N(R’)R”, R'NR 1 COR”, R'NR 1 C(O)OR”, R'NR 1 CONCR 2 )R ', R'NR 1 C(S)R ”, R’NR 1 S(O)R”, R’NR 1 S(O) 2 R ”, and R'NR 1 S(O) 2 N(R 2 )R ”, wherein R and R are independently selected from null, optionally substituted
  • R r is selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C 3 -C 13 fused cycloalkyl, optionally substituted C 3 -C 13 fused heterocyclyl, optionally substituted C 3 -C 13 bridged cycloalkyl, optionally substituted C 3 -C 13 bridged heterocyclyl, optionally substituted C 3 -C 13 spiro cycloalkyl, optionally substituted C 3 -C 13 spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R 1 and R 2 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C 2 -C 8 alkynyl, optionally substituted C 1 -C 8 alkoxyalkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 -C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R 1 , R 2 , R 3 and R 4 are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C 2 -C 8 alkynyl, optionally substituted C 1 -C 8 alkoxy, optionally substituted C 1 -C 8 alkoxyalkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 -C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylamino, and optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclylamino, optionally substituted 4-8 membered heterocycl
  • R 1 and R 2 , R 3 and R 4 together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring;
  • A, W and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R’-R”, R'COR”, R'CO 2 R”, R'C(O)N(R 5 )R”, R’C(S)N(R 5 )R”, R'OR”, R'OC(O)R”, R’OC(O)OR’ , R'OCONR 5 R”, R'SR ”, R'SOR”, R'SO 2 R”, R'SO 2 N(R 5 )R”, R'N(R 5 )R”, R’NR 5 COR”, R'NR 5 C(O)OR”, R’NR 5 CON(R 6 )R”, R’NR 5 C(S)R”, R’NR 5 S(O)R”, R’NR 5 S(O) 2 R”, and R'NR 5 S(O) 2 N(R 6 )R", wherein R’ and R” are independently selected from null, optionally substituted R r -(C 1 -C 8 alkyl), or a moiety
  • R 5 and R 6 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C 2 -C 8 alkynyl, optionally substituted C 1 -C 8 alkoxyalkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 -C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R 1 and R 2 are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, and optionally substituted C 1 -C 8 alkyl, optionally substituted C 1 -C 8 alkoxy, optionally substituted C 1 -C 8 alkoxy C 1 -C 8 alkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 -C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylamino, C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclylamino, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or
  • R 1 and R 2 together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring;
  • a and B are independently selected from null, or bivalent moiety selected from R’-R ”, R'COR”, R'CO 2 R”, R’C(O)NR 3 R”, R’C(S)NR 3 R”, R’OR”, R'OC(O)R “, R’OC(O)OR”, R'OCON(R 3 )R”, R’SR”, R’SOR”, R'SO 2 R”, R'SO 2 N(R 3 )R”, R'N(R 3 )R”, R’NR 3 COR”, R’NR 3 C(O)OR”, R’NR 3 CON(R 4 )R, R’NR 3 C(S)R”, R’NR 3 S(O)R”, R’NR 3 S(O) 2 R”, and R'NR 3 S(O) 2 N(R 4 )R”, wherein R’ and R” are independently selected from null, optionally substituted R r -(C 1 -C 8 alkyl), or a moiety comprising of optionally substituted C 1
  • R r is selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C 3 -C 13 fused cycloalkyl, optionally substituted C 3 -C 13 fused heterocyclyl, optionally substituted C 3 -C 13 bridged cycloalkyl, optionally substituted C 3 -C 13 bridged heterocyclyl, optionally substituted C 3 -C 13 spiro cycloalkyl, optionally substituted C 3 -C 13 spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R 3 and R 4 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C 2 -C 8 alkynyl, optionally substituted C 1 -C 8 alkoxyalkyl, optionally substituted C 1 -C 8 haloalky
  • R’ and R together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m, at each occurrence, is 0 to 15; and n is 0 to 15. wherein X is selected from O, NH, and NR 7 ;
  • R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C 2 -C 8 alkynyl, optionally substituted C 1 -C 8 alkoxy, optionally substituted C 1 -C 8 alkoxy C 1 -C 8 alkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 -C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylamino, optionally substituted C 1 -C 8 alkylaminoC 1 -C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 4-10 membered heterocyclyl, optionally substituted
  • a and B are independently selected from null, or bivalent moiety selected from R -R”, R'COR ”, R'CO 2 R”, R’C(O)N(R 8 )R”, R'C(S)N(R 8 )R”, R OR”, R OC(O)R”, R OC(O)OR”, R OCON(R 8 )R", R'SR”, R’SOR ”, R'SO 2 R", R’SO 2 N(R 8 )R”, R’N(R 8 )R”, R’NR 8 COR”, R’NR 8 C(O)OR”, R'NR 8 CON(R 9 )R”, R'NR 8 C(S)R”, R’NR 8 S(O)R”, R'NR 8 S(O) 2 R”, and R’NR 8 S(O) 2 N(R 9 )R”, wherein R’ and R” are independently selected from null, optionally substituted R r -(C 1 -C 8 alkyl), or a moiety comprising of optionally substituted C
  • R r is selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C 3 -C 13 fused cycloalkyl, optionally substituted C 3 -C 13 fused heterocyclyl, optionally substituted C 3 -C 13 bridged cycloalkyl, optionally substituted C 3 -C 13 bridged heterocyclyl, optionally substituted C 3 -C 13 spiro cycloalkyl, optionally substituted C3-C 13 spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R 7 , R 8 and R 9 are independently selected from hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted C 2 -C 8 alkenyl, optionally substituted C2-C 8 alkynyl, optionally substituted C 1 -C 8 alkoxyalkyl, optionally substituted C 1 -C 8 haloalkyl, optionally substituted C 1 - C 8 hydroxyalkyl, optionally substituted C 1 -C 8 alkylaminoCi-C 8 alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
  • R’ and R”, R 8 and R 9 , R’ and R 8 , R’ andR 9 , R” and R 8 , R” and R 9 together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m, at each occurrence, is 0 to 15; n, at each occurrence, is 0 to 15; o is 0 to 15; and p is 0 to 15.
  • the linker moiety comprises a ring selected from the group consisting of a 3 to 13 membered ring, a 3 to 13 membered fused ring, a 3 to 13 membered bridged ring, and a 3 to 13 membered spiro ring.
  • the linker moiety comprises a ring selected from the group consisting of FORMULA C1, C2, C3, C4 and C5:
  • R has a structure of:
  • the G9a/GLP degrader is compound selected from the following compounds, as identified in Table 1 below: YH72-103, YH72-104, YH72-106, YH72-136, YH72- 137, YH72-138, YH72-139, YH72-140, YH72-141, YH72-142, YH72-143, YH72-144, YH72-
  • novel synthesized bivalent compounds i.e., G9a/GLP degraders/disruptors
  • G9a/GLP degraders/disruptors can be assessed using standard biophysical assays known in the art (e.g., isothermal titration calorimetry (ITC), surface plasmon resonance (SPR)).
  • Cellular assays can then be used to assess the bivalent compound’s ability to induce G9a/GLP degradation and inhibit cancer cell proliferation.
  • enzymatic activity can also be assessed.
  • Assays suitable for use in any or all of these steps are known in the art, and include, e.g., Western blotting, quantitative mass spectrometry (MS) analysis, flow cytometry, enzymatic inhibition, ITC, SPR, cell growth inhibition and xenograft and PDX models.
  • MS mass spectrometry
  • Suitable cell lines for use in any or all of these steps are known in the art and include, cancer cell lines: 1) Prostate (22Rv1, PC-3, DU145); 2) Leukemia (K562, THP-1, A9M); 3) Lung (H1299, A549, H1975, PC 14, H441, CL 1- 5); 4) Breast (MDA-MB231, MCF7, MCF10A, SKBR-3, MDA-MB435, ZR-75-30); 5) Ovarian (DOV-13, HeyC2, OV2008, 0vCAR5); 6) Pancreatic (PANC-1, BxP3, Capanl, Hs766T, MIAPaCa2); 7) bladder (T24, J82, RT112, 5637).
  • Suitable mouse models for use in any or all of these steps are known in the art and include patient-derived xenograft models of triple negative breast cancer.
  • isotopic variations of the compounds disclosed herein are contemplated and can be synthesized using conventional methods known in the art or methods corresponding to those described in the Examples (substituting appropriate reagents with appropriate isotopic variations of those reagents).
  • an isotopic variation is a compound in which at least one atom is replaced by an atom having the same atomic number, but an atomic mass different from the atomic mass usually found in nature.
  • Useful isotopes are known in the art and include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine. Exemplary isotopes thus include, e.g., 2 H, 3 H, 13 C, 1 1 4 4 .C, 15 N, 17 O, 18 O, 32 P, 35 S, 18 F, and 36 C1.
  • a fluorinated variation is a compound in which at least one hydrogen atom is replaced by a fluoro atom. Fluorinated variations can provide therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements.
  • prodrugs of the compounds disclosed herein are contemplated and can be synthesized using conventional methods known in the art or methods corresponding to those described in the Examples (e.g., converting hydroxyl groups or carboxylic acid groups to ester groups).
  • a prodrug refers to a compound that can be converted via some chemical or physiological process (e.g., enzymatic processes and metabolic hydrolysis) to a therapeutic agent.
  • prodrug also refers to a precursor of a biologically active compound that is pharmaceutically acceptable.
  • a prodrug may be inactive when administered to a subject, i.e.
  • prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in an organism.
  • prodrug is also meant to include any covalently bonded carriers, which release the active compound in vivo when such prodrug is administered to a subject.
  • Prodrugs of an active compound may be prepared by modifying functional groups present in the active compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent active compound.
  • Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of an alcohol or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound and the like.
  • Figure 1 The effect of Examples 29, 30, 31, 35, 37,39, 41, and 44 on reducing the G9a and GLP protein levels.
  • MS8709 degrades G9a and GLP and inhibits cell growth in K562 cells.
  • FIG 12D Representative WB analysis of G9a and GLP protein levels in 22Rv1 cells treated with compound 5 or 9 at the indicated concentration for 24 h. Vinculin and ⁇ -actin were used at loading controls.
  • Figure 12E Clonogenicity assay of 22Rv1 cells following 8 d treatment with 5, UNC0642, and 9 at indicated concentrations. Results shown are representative of three independent biological experiments.
  • Figure 12F WB analysis of G9a and GLP protein levels in 22Rv1 cells pre-treated with UNC0642 (10 ⁇ M), MG132 (5 ⁇ M), MLN4942 (1 ⁇ M) or VHL-1 (5 ⁇ M) for 2 h and subsequently treated with compound 5 (1 ⁇ M) for 24 h. Vinculin and ⁇ -actin were used as loading controls.
  • WB results shown in panels D and F are representative of two independent biological experiments.
  • Alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation.
  • An alkyl may comprise one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms.
  • an alkyl comprises one to fifteen carbon atoms (e.g., C 1 -C 15 alkyl).
  • an alkyl comprises one to thirteen carbon atoms (e.g, C 1 -C 13 alkyl).
  • an alkyl comprises one to eight carbon atoms (e.g, C 1 -C 8 alkyl).
  • Alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond.
  • An alkenyl may comprise two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms.
  • an alkenyl comprises two to twelve carbon atoms (e.g., C 2 -C 12 alkenyl).
  • an alkenyl comprises two to eight carbon atoms (e.g., C 2 -C 8 alkenyl).
  • an alkenyl comprises two to six carbon atoms (e.g., C 2 - C 6 alkenyl).
  • an alkenyl comprises two to four carbon atoms (e.g., C 2 -C 4 alkenyl).
  • the alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, penta- 1,4-dienyl, and the like.
  • alkynyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one triple bond.
  • An alkynyl may comprise two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms.
  • an alkynyl comprises two to twelve carbon atoms (e.g., C 2 -C 12 alkynyl).
  • an alkynyl comprises two to eight carbon atoms (e.g., C 2 -C 8 alkynyl).
  • an alkynyl has two to six carbon atoms (e.g., C 2 -C 6 alkynyl). In other embodiments, an alkynyl has two to four carbon atoms (e.g., C 2 -C 4 alkynyl).
  • the alkynyl is attached to the rest of the molecule by a single bond. Examples of such groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, 1 -pentynyl, 2-pentynyl, 1 -hexynyl, 2-hexynyl, 3 -hexynyl, and the like.
  • alkoxy means an alkyl group as defined herein witch is attached to the rest of the molecule via an oxygen atom.
  • alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, iso-propyloxy, n-butoxy, iso-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like.
  • aryl refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom.
  • the aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and carbon atoms.
  • An aryl may comprise from six to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) ⁇ -electron system in accordance with the Huckel theory.
  • an aryl comprises six to fourteen carbon atoms (C 6 -C 14 aryl).
  • an aryl comprises six to ten carbon atoms (C 6 -C 10 aryl).
  • groups include, but are not limited to, phenyl, fluorenyl and naphthyl.
  • Heteroaryl includes fused or bridged ring systems
  • a heteroaryl refers to a radical derived from a 3- to 10-membered aromatic ring radical (3-10 membered heteroaryl).
  • a heteroaryl refers to a radical derived from 5- to 7-membered aromatic ring (5-7 membered heteroaryl).
  • Heteroaryl includes fused or bridged ring systems.
  • the heteroatom(s) in the heteroaryl radical is optionally oxidized.
  • One or more nitrogen atoms, if present, are optionally quatemized.
  • the heteroaryl is attached to the rest of the molecule through any atom of the ring(s).
  • Examples of such groups include, but not limited to, pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazoliny
  • heterocyclyl means a non-aromatic, monocyclic, bicyclic, tricyclic, or tetracyclic radical having a total of from 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 atoms in its ring system, and containing from 3 to 12 carbon atoms and from 1 to 4 heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms.
  • a heterocyclyl group may include fused, bridged or spirocyclic ring systems. In certain embodiments, a heterocyclyl group comprises 3 to 10 ring atoms (3-10 membered heterocyclyl).
  • a heterocyclyl group comprises 3 to 8 ring atoms (3-8 membered heterocyclyl). In certain embodiments, a heterocyclyl group comprises 4 to 8 ring atoms (4-8 membered heterocyclyl). In certain embodiments, a heterocyclyl group comprises 3 to 6 ring atoms (3-6 membered heterocyclyl).
  • a heterocyclyl group may contain an oxo substituent at any available atom that will result in a stable compound. For example, such a group may contain an oxo atom at an available carbon or nitrogen atom. Such a group may contain more than one oxo substituent if chemically feasible.
  • heterocyclyl group when such a heterocyclyl group contains a sulfur atom, said sulfur atom may be oxidized with one or two oxygen atoms to afford either a sulfoxide or sulfone.
  • An example of a 4 membered heterocyclyl group is azetidinyl (derived from azetidine).
  • An example of a 5 membered cycloheteroalkyl group is pyrrolidinyl.
  • An example of a 6 membered cycloheteroalkyl group is piperidinyl.
  • An example of a 9 membered cycloheteroalkyl group is indolinyl.
  • An example of a 10 membered cycloheteroalkyl group is 4H-quinolizinyl.
  • Such heterocyclyl groups include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3- pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, di
  • cycloalkyl means a saturated, monocyclic, bicyclic, tricyclic, or tetracyclic radical having a total of from 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 carbon atoms in its ring system.
  • a cycloalkyl may be fused, bridged or spirocyclic.
  • a cycloalkyl comprises 3 to 8 carbon ring atoms (C 3 -C 8 cycloalkyl).
  • a cycloalkyl comprises 3 to 6 carbon ring atoms (C 3 -C 6 cycloalkyl).
  • Examples of such groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, adamantyl, and the like.
  • cycloalkylene is a bidentate radical obtained by removing a hydrogen atom from a cycloalkyl ring as defined above.
  • groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, cyclopentenylene, cyclohexylene, cycloheptylene, and the like.
  • spirocyclic as used herein has its conventional meaning, that is, any ring system containing two or more rings wherein two of the rings have one ring carbon in common.
  • Each ring of the spirocyclic ring system independently comprises 3 to 20 ring atoms. Preferably, they have 3 to 10 ring atoms.
  • Non-limiting examples of a spirocyclic system include spiro[3.3]heptane, spiro[3.4] octane, and spiro[4.5]decane.
  • cyano refers to a -ON group.
  • aldehyde refers to a -C(O)H group.
  • alkoxy refers to both an -O-alkyl, as defined herein.
  • alkoxycarbonyl refers to a -C(O)-alkoxy, as defined herein.
  • alkylaminoalkyl refers to an -alkyl-NR-alkyl group, as defined herein.
  • alkylsulfonyl refer to a -SO 2 alkyl, as defined herein.
  • amino refers to an optionally substituted -NH 2 .
  • aminoalkyl refers to an -alky-amino group, as defined herein.
  • aryloxy refers to both an -O-aryl and an -O-heteroaryl group, as defined herein.
  • aryloxycarbonyl refers to -C(O)-aryloxy, as defined herein.
  • aryl sulfonyl refers to a -SO 2 aryl, as defined herein.
  • carbonyl refers to a -C(O)- group, as defined herein.
  • a “carboxylic acid” group refers to a -C(O)OH group.
  • cycloalkoxy refers to a -O-cycloalkyl group, as defined herein.
  • halo or halogen group refers to fluorine, chlorine, bromine or iodine.
  • haloalkyl group refers to an alkyl group substituted with one or more halogen atoms.
  • a "hydroxy” group refers to an -OH group.
  • a "nitro” group refers to a -NO 2 group.
  • the same symbol in a different FORMULA may have a different definition, for example, the definition of R 1 in FORMULA 1 is different from that in FORMULA 3 A.
  • pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d- ⁇ -tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes,
  • Particularly favored derivatives and prodrugs are those that increase the bioavailability of the compounds disclosed herein when such compounds are administered to a mammal (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species.
  • Preferred prodrugs include derivatives where a group that enhances aqueous solubility or active transport through the gut membrane is appended to the structure of FORMULAE described herein. Such derivatives are recognizable to those skilled in the art without undue experimentation. Nevertheless, reference is made to the teaching of Burger’s Medicinal Chemistry and Drug Discovery, 5 th Edition, Vol. 1: Principles and Practice, which is incorporated herein by reference to the extent of teaching such derivatives.
  • Salts derived from appropriate bases include, e.g., alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N-(alkyl)4+ salts.
  • alkali metal e.g., sodium
  • alkaline earth metal e.g., magnesium
  • ammonium e.g., ammonium
  • N-(alkyl)4+ salts e.g., sodium
  • alkaline earth metal e.g., magnesium
  • ammonium e.g., sodium
  • N-(alkyl)4+ salts e.g., sodium
  • alkali metal e.g., sodium
  • alkaline earth metal e.g., magnesium
  • ammonium e.g., sodium
  • N-(alkyl)4+ salts e.g., ammonium and N-(alkyl)4+ salts.
  • the invention also envisions the quaternization of any basic nitrogen-
  • parenteral includes subcutaneous, intracutaneous, intravenous, intramuscular, intraperitoneal, intra-articular, intra-arterial, intrasynovial, intrastemal, intrathecal, intralesional, and intracranial injection or infusion techniques.
  • compositions of this invention can be administered in the form of suppositories for rectal administration.
  • These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components.
  • suitable non-irritating excipient include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.
  • any bland fixed oil can be employed, including synthetic mono- or diglycerides.
  • Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically- acceptable oils, e.g., olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions.
  • Other commonly used surfactants such as Tweens, Spans, or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms can also be used for the purposes of formulation.
  • multiple parties can be included in subject selection.
  • a first party can obtain a sample from a candidate subject and a second party can test the sample.
  • subjects can be selected or referred by a medical practitioner (e.g., a general practitioner).
  • subject selection can include obtaining a sample from a selected subject and storing the sample or using the in the methods disclosed herein. Samples can include, e.g., cells or populations of cells.
  • the term “treating cancer’ ’ means causing a partial or complete decrease in the rate of growth of a tumor, and/or in the size of the tumor and/or in the rate of local or distant tumor metastasis, and/or the overall tumor burden in a subject, and/or any decrease in tumor survival, in the presence of a degrader/disruptor (e.g., an G9a/GLP degrader/ disruptor) described herein.
  • a degrader/disruptor e.g., an G9a/GLP degrader/ disruptor
  • prevent shall refer to a decrease in the occurrence of a disease or decrease in the risk of acquiring a disease or its associated symptoms in a subject.
  • the prevention may be complete, e.g., the total absence of disease or pathological cells in a subject.
  • the prevention may also be partial, such that the occurrence of the disease or pathological cells in a subject is less than, occurs later than, or develops more slowly than that which would have occurred without the present invention.
  • Exemplary G9a/GLP- mediated diseases that can be treated with G9a/GLP degraders/ disruptors include, for example, breast cancer, ovarian cancer, prostate cancer, colon cancer, pancreatic cancer, bladder cancer, liver cancer and cervical cancer.
  • the subject can be evaluated to detect, assess, or determine their level of disease.
  • treatment can continue until a change (e.g., reduction) in the level of disease in the subject is detected.
  • a maintenance dose of a compound, or composition disclosed herein can be administered, if necessary.
  • the dosage or frequency of administration, or both can be reduced, e.g., as a function of the symptoms, to a level at which the improved condition is retained.
  • Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.
  • the compound I-3 (212.8 mg, 0.4 mmol) was dissolved in i-PrOH (3 mL) and treated with N, N-diisopropylethylamine (0.2 mL, 1.2 mmol) and 4,4-difluoropiperidine (187.5 mg, 1.2 mmol). The solution was heated in a microwave reactor at 130 °C for 30 min before being concentrated under reduced pressure.
  • Example 1 (N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-2-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yI)amino)acetamide) (YH72-103) HOAt (3.8 mg, 0.027 mmol, 1.5 equiv), EDCI (5.4 mg, 0.027 mmol, 1.5 equiv) and NMM (7.1 mg, 0.054 mmol, 3.0 equiv) were added to a solution ofI-5 (11.8 mg, 0.018mmol) and L-l (6.5 mg, 1.0 equiv) in DMSO (1 mL).
  • Example 2- 28 were synthesized following the same procedure for preparing Example 1.
  • Example 2 ( N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-4-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yl)amino)butanamide) (YH72-104) 52% yield.
  • Example 3 ( N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-4-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yl)amino)butanamide) (YH72-106) 52% yield.
  • Example 5 (N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-6-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yl)amino)hexanamide) (YH72-137) 64% yield.
  • Example 6 (N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-7-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yl)amino)heptanamide) (YH72-138) 56% yield.
  • Example 7 ( N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-8-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisoindolin-4-yl)amino)octanamide) (YH72-139) 47% yield.
  • Example 9 (N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-3-(2-(2-((2-(2,6-dioxopiperidin-3- yl)-l,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)pr open amide) (YH72-141) 58% yield.
  • Example 10 ( N-(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-3-(2-(2-(2-((2-(2,6-dioxopiperidin- 3-yl)-l,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propenamide) (YH72-142) 42% yield.
  • Example 13 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 4 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)succinimide) (YH72-122) 39% yield.
  • Example 14 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 5 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)glutaramide) (YH72-119) 62% yield.
  • Example 15 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 6 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)adipamide) (YH72-114) 57% yield.
  • Example 16 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 7 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)heptanediamide) (YH72-109) 68% yield.
  • Example 17 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 8 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)octanediamide) (YH72-135) 53% yield.
  • Example 18 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 9 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)nonanediamide) (YH72-108) 48% yield.
  • Example 19 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 10 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- y l)d ecan ediam ide) (YH72-107) 42% yield.
  • Example 20 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 11 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)undecanediamide) (YH72-134) 41% yield.
  • Example 21 ((2S,4R)-1-((S)-2-(2-(2-((3-(4-((2-(4,4-difluoropiperidin-1-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)propyl)amino)-2-oxoethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide) (YH72-115) 56% yield.
  • Example 2222 : ((2S,4R)-1-((S)-2-(3-(3-((3-(4-((2-(4,4-difluoropiperidin-1-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)propyl)amino)-3-oxopropoxy)propanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide) (YH72-116) 38% yield.
  • Example 23 ((2S, 4R)- l-((S)-2-(tert-butyl)- 15-(4-((2-(4,4-difluoropiperidin- l-yl)-6- methoxy-7-(3-(pyrrolidin- l-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-4, 11-dioxo- 6,9-dioxa-3,12-diazapentadecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH72-117) 65% yield.
  • Example 24 ((2S,4R)- l-((S)-2-(tert-butyl)- 17-(4-((2-(4,4-difluoropiperidin- l-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-4,13-dioxo- 7, 10-dioxa-3, 14-diazaheptadecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH72-118) 58% yield.
  • Example 25 ((2S,4R)-1-((S)-2-(tert-butyl)-20-(4-((2-(4,4-difluoropiperidin-1-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-4,16-dioxo- 7,10,13-trioxa-3,17-diazaicosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH72-120) 86% yield.
  • Example 26 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 16 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)- 4,7,10,13-tetraoxahexadecanediamide) (YH72-121) 13% yield.
  • Example 27 (N 1 -(3-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)propyl)-N 17 -((S)-1-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)- 3,6,9,12,15-pentaoxaheptadecanediamide) (YH72-133) 44% yield.
  • Example 29 ((2S,4R)-1-((S)-2-(3-(2-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy- 7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)ethoxy)propanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide) (YH80-79) HATU (5.1 mg, 0.012 mmol, 1.0 equiv) and DIPEA (20 ⁇ L, 0.24 mmol, 20 equiv) were added to a solution of I-8 (10.4 mg, 0.012 mmol) and L-29 (9.7 mg, 1.0 equiv) in DMSO (1 mL).
  • Example 31 ((2S,4R)-1-((S)-2-(tert-butyl)-20-(4-((2-(4,4-difluoropiperidin-1-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-4,17-dioxo- 7,10,13-trioxa-3,16-diazaicosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH80-94) 66% yield.
  • Example 36 ((2S,4R)-1-((S)-2-(4-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)butanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH80-99) 62% yield).
  • Example 40 ((2S,4R)-1-((S)-2-(8-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yI)butanamido)octanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH80-103) 29% yield.
  • Example 41 ((2S,4R)-1-((S)-2-(9-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)nonanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH80-104) 61% yield.
  • Example 42 ((2S,4R)-1-((S)-2-(10-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)decanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH80-105) 74% yield.
  • Example 45 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)ethyl)butanamide (YH80-39) 41% yield.
  • Example 47 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(4-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)butyl)butanamide (YH80-41) 38% yield.
  • Example 48 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(5-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)pentyl)butanamide (YH80-42) 50% yield.
  • Example 49 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(6-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)hexyl)butanamide (YH 80-43) 47% yield.
  • Example 50 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(7-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)heptyl)butanamide (YH80-44) 46% yield.
  • Example 52 (4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(10-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)decyl)butanamide) (YH87-016) 57% yield.
  • Example 56 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)- l,3-dioxoisomdolin-4-yl)ammo)ethoxy)ethoxy)ethyl)butanamide (YH80-35) 48% yield.
  • Example 57 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3- yl)-l,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl) butanamide (YH80-36) 42% yield.
  • Example 58 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(14-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxatetradecyl) butanamide (YH80-37) 30% yield.
  • Example 59 4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-N-(17-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaheptadecyl)butanamide (YH80-38) 44% yield.
  • Examples 60-87 were synthesized following similar procedure for preparing example 29.
  • Example 60 ((2S,4R)-.N-((S)-3-((2-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)ethyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocydopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-042) 72% yield.
  • Example 61 ((2S,4R)-N-((S)-3-((3-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)propyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-043) 80% yield.
  • Example 62 ((2S,4R)-N-((S)-3-((4-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)butyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-044) 87% yield.
  • Example 63 ((2S,4R)-N-((S)-3-((5-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)pentyI)amino)-1-(4-(4-methyIthiazol-5-yl)phenyI)-3-oxopropyI)-1-(2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-045) 84% yield.
  • Example 64 ((2S,4R)-N-((S)-3-((6-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy- 7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)hexyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-046) 97% yield.
  • Example 65 ((2S,4R)-N-((S)-3-((7-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)heptyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-047) 72% yield.
  • Example 66 ((2S,4R)-N-((S)-3-((8-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)octyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-048) 79% yield.
  • Example 67 ((2S,4R)-N-((S)-3-((9-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)nonyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-049) 87% yield.
  • Example 68 ((2S,4R)-N-((S)-3-((10-(4-(4-((2-(4,4-difluoropiperidiii-1-yl)-6-methoxy- 7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)decyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyI)-4-hydroxypyrrolidine-2- carboxamide) (YH87-090) 68% yield.
  • Example 69 ((2S,4R)-N-((S)-3-((l l-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy- 7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)undecyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-050) 68% yield.
  • Example 70 ((2S,4R)-N-((S)-3-((12-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy- 7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)dodecyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide) (YH99-051) 76% yield.
  • Example 7711 (2S,4R)-N-((S)-3-((2-(2-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6- methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)ethoxy)ethyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)- 2-(l-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide (YH119-13) 60% yield.
  • Example 72 (2S,4R)-N-((S)-17-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3- (pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-1-(4-(4-methylthiazol-5- yl)phenyl)-3,14-dioxo-7,10-dioxa-4,13-diazaheptadecyl)-1-((S)-2-(l-fluorocyclopropane-1- carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (YH119-14) 90% yield.
  • Example 73 (2S,4R)-N-((S)-20-(4-((2-(4,4-difluoropiperidin-1-yI)-6-methoxy-7-(3- (pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-1-(4-(4-methylthiazol-5- yl)phenyl)-3,17-dioxo-7,10,13-trioxa-4,16-diazaicosyl)-1-((S)-2-(l-fluorocyclopropane-1- carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (YH119-15) 96% yield.
  • Example 74 (2S,4R)-N-((S)-23-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3- (pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-1-(4-(4-methylthiazol-5- yl)phenyl)-3,20-dioxo-7, 10,13, 16-tetraoxa-4, 19-diazatricosyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide (YH119-16) 81% yield.
  • Example 75 (2S,4R)-N-((S)-26-(4-((2-(4,4-difluoropiperidin-1-yI)-6-methoxy-7-(3- (pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1-yl)-1-(4-(4-methylthiazol-5- yl)phenyl)-3,23-dioxo-7, 10, 13,16, 19-pentaoxa-4, 22-diazahexacosyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide (YH119-17) 80% yield.
  • Example 76 (2S,4R)-N-(2-(2-((2-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)ethyl)amino)-2-oxoethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(l- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide (YH119-001) 98% yield.
  • Example 83 (2S,4R)-N-(2-(2-((2-(2-(4-(4-((2-(4,4-diHuoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)ethoxy)ethyl)amino)-2-oxoethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2- (l-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamide (YH119-008) 65% yield.
  • Example 8866 (2S,4R)-N-(2-((22-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-
  • Examples 88-94 were synthesized following similar procedure for preparing example 29.
  • Example 88 ((2S,4R)-1-((S)-2-(12-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrroIidin-1-yI)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)dodecanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(l-(4-(4-methylthiazol- 5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide) (YH87-171) 59% yield.
  • Example 90 N-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decyl)- 4-(4-((6-methoxy-2-morpholino-7-(3-(pyrrolidin-1-yl)propoxy)qiiinolin-4- yl)amino)piperidin-1-yl)butanamide (YH87-174) 58% yield.
  • MS (ESI) [M+H] + 966.5.
  • Example 91 (N-(ll-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)undecyl)-4-(4-((6-methoxy-2-morpholino-7-(3-(pyrrolidin-1-yl)propoxy)quinolin- 4-yl)amino)piperidin-1-yl)butanamide) (YH87-175) 36% yield.
  • Example 92 (N-(12-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)dodecyl)-4-(4-((6-methoxy-2-morpholino-7-(3-(pyrrolidin-1-yl)propoxy)quinolin- 4-yl)amino)piperidin-1-yl)butanamide) (YH87-176) 39% yield.
  • Example 93 ((2R,4S)-1-((S)-2-(12-(4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7- (3-(pyrrolidin-1-yl)propoxy)quinazolin-4-yl)amino)piperidin-1- yl)butanamido)dodecanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide) (YH87-183) 90% yield.
  • Example 9944 (4-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methoxy-7-(3-(pyrrolidin-1- yl)propoxy)quinolin-4-yl)amino)piperidin-1-yl)-N-(12-((2-(l-methyl-2,6-dioxopiperidin-3- yl)-l,3-dioxoisoindolin-4-yl)amino)dodecyl)butanamide) (YH87-184) 94% yield.
  • Example compounds are set forth in Table 1 below.
  • the structure shall control.
  • UNC0642 is a highly potent G9a/GLP inhibitor (IC 50 ⁇ 2.5 nM for G9a and GLP in biochemical assays). 42 Second, UNC0642 is highly selective, showed minimal off-target toxicity, and is suitable for in vivo efficacy studies. 42 Third, we previously solved the cocrystal structure of G9a in complex with UNC0638, a close analog of UNC0642 (PDB: 3RJW).
  • Compound 5 induces G9a/GLP degradation in a concentration- and time-dependent manner and inhibits 22Rvl cell growth.
  • Compound 5 but not its parent inhibitor UNC0642, potently induced degradation of G9a and GLP at 1 ⁇ M following 24 h treatment (Figure 11A). Consistent with its G9a/GLP degradation activity, compound 5 inhibited 22Rv1 cell growth after 7 d treatment with an Gl 50 of 4.1 ⁇ M, while UNC0642 did not ( Figure 11B). This data highlights the superior antiproliferative activity of G9a/GLP PROTAC degraders to G9a/GLP catalytic inhibitors.
  • compound 5 potently degrades G9a and GLP in a concentration- and time-dependent manner and has more effective anti-proliferative activity in 22Rv1 cells compared to the parent G9a/GLP catalytic inhibitor UNC0642.
  • the G9a/GLP degradation induced by compound 5 is dependent on VHL and UPS.
  • Cpds 2-8 were synthesized following the same procedure for preparing cpd 1.
  • Example 114 The effect of Examples 29, 30, 31, 35, 37, 39, 4,1 and 44 (MS8709), on reducing the G9a and GLP protein levels.
  • 22Rv1 cells were treated with DMSO or the indicated compound at 0.3 or 3 ⁇ M for 24 h.
  • the cell lysates were analyzed via WB to examine the G9a and GLP protein levels with vinculin as the loading control. The percent of protein remaining was determined by normalizing to DMSO controls.
  • Example 115 Characterization of MS8709 in 22Rv1 cells.
  • MS8709 induces G9a/GLP degradation in a VHL- and UPS-dependent manner and MS8709, but not UNC0642 or its negative control (Example 93), suppresses clonogenicity in 22Rv1 cells.
  • B-C G9a (B) and GLP (C) enzymatic activity of MS8709, UNC0642, and Example 93 at indicated concentrations in G9a and GLP radioactive methyltransferase assays.
  • MS8709 is selective for G9a and GLP over other protein methyltransferases. Selectivity of MS8709 (at 10 ⁇ M) against a panel of 21 protein methyltransferases was assessed using radioactive methyltransferase assays.
  • Example 118 MS8709 degrades G9a and GLP and inhibits cell growth in K562 cells.
  • Example 119 MS8709 degrades G9a and GLP and inhibits proliferation in H1299 cells.
  • Example 120 Plasma concentrations of MS8709 over an 8-h period in mice after a single IP injection of MS8709 at 50 mg/kg.
  • 22Rv1, H1299, and K562 cells were purchased from ATCC. 22Rv1 and H1299 were cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin-streptomycin. K562 cells were grown in IMDM containing 10% FBS and 1% penicillin-streptomycin. All cells were cultured at 37° C with 5% CO 2 .
  • 22Rv1, K562, and H1299 cells were seeded in 96 well-plates and treated with compound at indicated serial dilations for 7-8 days. Plates were incubated at 37° C with 5% CO 2 . Cell viability was determined using CCK-8 (Cell Counting Kit-8, Dojindo, CK04). Absorbance and reference values were measured at 450 nm and 690 nm respectively on the Infinite F PLEX plate reader (TEC AN). IC 50 values were calculated using GraphPad Prism 6. Clonogenicity assay
  • 22Rv1 cells were seeded at 50,000 cells per well in 6 well plates and treated continuously with compounds at indicated concentrations for 8 days. After 8 days, the cells were stained with 0.5% crystal violet dye.
  • G9a and GLP enzymatic activity assays were conducted in duplicate at indicated concentrations using the radioisotope-based MT HotSpotTM system by Reaction Biology Corp.
  • the selectivity panel of 21 methyltransferases was performed in duplicate with the same assay by Reaction Biology Corp, using 10 ⁇ M of compound.
  • RT-qPCR was performed according to a previously described protocol 31 (Cao et al., 2014). H1299 cells were treated with DMSO, UNC0642 (3 ⁇ M), or MS8709 (3 ⁇ M) for 24 h. Total RNA was extracted using the Monarch Total RNA Miniprep Kit (T2010S, New England Biolabs) and cDNA was synthesized using the SuperScript IV First-Strand Synthesis System (18091050, Thermofisher). qPCR was performed on the Agilent Technologies Stratagene Mx3005p qPCR system. Primer sets used are listed below.
  • Histone H3 lysine 9 methyltransferase G9a is a transcriptional coactivator for nuclear receptors. J. Biol. Chem. 281, 8476-8485. 10.1074/jbc.M511093200. 25. Lehnertz, B., Pabst, C., Su, L., Miller, M., Liu, F., Yi, L., Zhang, R., Krosl, J., Yung, E., Kirschner, J., et al. (2014). The methyltransferase G9a regulates HoxA9-dependent transcription in AML. Genes Dev. 28, 317-327. 10.1101/gad.236794.113.
  • Homo-PROTACs bivalent small-molecule dimerizers of the VHL E3 ubiquitin ligase to induce self-degradation. Nat Commun 8, 830. 10.1038/s41467-017- 00954-1.
  • IAP antagonists induce autoubiquitination of c-IAPs, NF-kappaB activation, and TNF alpha-dependent apoptosis.
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Abstract

L'invention divulgue des composés dégradant/perturbant G9a/GLP qui se lient à G9a/GLP et et induisent leur dégradation, éliminant ainsi toutes les fonctions d'échafaudage de G9a/GLP en plus de l'activité kinase de G9a/GLP. Les agents de dégradation de G9a/GLP divulgués dans la description sont des composés bivalents, comprenant un ligand G9a/GLP conjugué à une étiquette de dégradation/perturbation par l'intermédiaire d'un lieur.
PCT/US2025/012138 2024-01-19 2025-01-17 Composés dégradant/perturbant g9a/glp et procédés d'utilisation Pending WO2025155882A1 (fr)

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Citations (2)

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WO2023064586A1 (fr) * 2021-10-15 2023-04-20 Tango Therapeutics, Inc. Nouveaux modulateurs de l'ehmt1 et de l'ehmt2 et leur utilisation thérapeutique
WO2024000763A1 (fr) * 2022-06-27 2024-01-04 中山大学 Inhibiteur covalent g9a/glp, son procédé de préparation et son utilisation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023064586A1 (fr) * 2021-10-15 2023-04-20 Tango Therapeutics, Inc. Nouveaux modulateurs de l'ehmt1 et de l'ehmt2 et leur utilisation thérapeutique
WO2024000763A1 (fr) * 2022-06-27 2024-01-04 中山大学 Inhibiteur covalent g9a/glp, son procédé de préparation et son utilisation

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VELEZ JULIA, HAN YULIN, YIM HYERIN, YANG PEIYI, DENG ZHIJIE, PARK KWANG-SU, KABIR MD, KANISKAN H. ÜMIT, XIONG YAN, JIN JIAN: "Discovery of the First-in-Class G9a/GLP PROTAC Degrader", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 67, no. 8, 25 April 2024 (2024-04-25), US , pages 6397 - 6409, XP093338759, ISSN: 0022-2623, DOI: 10.1021/acs.jmedchem.3c02394 *

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